Marie-Pierre Chauzat

A Survey of Pesticide Residues in Pollen Loads Collected by Honey Bees in France

In 2002, a field survey was initiated on French apiaries to monitor weakness of honey bee, Apis mellifera L., colonies. Apiaries were evenly distributed in five sites located on continental France. Five colonies were randomly selected in each apiary, leading to a total of 125 studied honey bee colonies. For 3 yr (starting in autumn 2002), colonies were visited four times per year: after winter, before summer, during summer, and before winter. Pollen loads from traps were collected at each visit. Multiresidue analyses were performed in pollen to search residues of 36 different molecules. Specific analyses were conducted to search fipronil and metabolites and also imidacloprid and metabolites. Residues of 19 searched compounds were found in samples. Contamination by pesticides ranged from 50 to 0%. Coumaphos and tau-fluvalinate residues were the most concentrated of all residues (mean concentrations were 925.0 and 487.2 microg/kg, respectively). Fipronil and metabolite contents were superior to the limit of detection in 16 samples. Residues of fipronil were found in 10 samples. Nine samples contained the sulfone compound, and three samples contained the desulfinyl compound. Residues of imidacloprid and 6-chloronicotinic acid were found in 69% of samples. Imidacloprid contents were quantified in 11 samples with values ranging from 1.1 to 5.7 microg/kg. 6-Chloronicotinic acid content was superior to the limit of quantification in 28 samples with values ranging from 0.6 to 9.3 microg/kg. Statistical tests showed no difference between places of sampling with the exception of fipronil. Possible origins of these contaminations, concentration and toxicity of pesticides, and the possible consequences for bees are discussed.

Standard methods for Nosema research

Summary Methods are described for working with Nosema apis and Nosema ceranae in the field and in the laboratory. For fieldwork, different sampling methods are described to determine colony level infections at a given point in time, but also for following the temporal infection dynamics. Suggestions are made for how to standardise field trials for evaluating treatments and disease impact. The laboratory methods described include different means for determining colony level and individual bee infection levels and methods for species determination, including light microscopy, electron microscopy, and molecular methods (PCR). Suggestions are made for how to standardise cage trials, and different inoculation methods for infecting bees are described, including control methods for spore viability. A cell culture system for in vitro rearing of Nosema spp. is described. Finally, how to conduct different types of experiments are described, including infectious dose, dose effects, course of infection and longevity tests.

Influence of Pesticide Residues on Honey Bee (Hymenoptera: Apidae) Colony Health in France

ABSTRACT A 3-yr field survey was carried out in France, from 2002 to 2005, to study honey bee (Apis mellifera L.) colony health in relation to pesticide residues found in the colonies. This study was motivated by recent massive losses of honey bee colonies, and our objective was to examine the possible relationship between low levels of pesticide residues in apicultural matrices (honey, pollen collected by honey bees, beeswax) and colony health as measured by colony mortality and adult and brood population abundance. When all apicultural matrices were pooled together, the number of pesticide residue detected per sampling period (four sampling periods per year) and per apiary ranged from 0 to 9, with the most frequent being two (29.6%). No pesticide residues were detected during 12.7% of the sampling periods. Residues of imidacloprid and 6- chloronicotinic acid were the most frequently detected in pollen loads, honey, and honey bee matrices. Several pairs of active ingredients were present concurrently within honey bees and in pollen loads but not in beeswax and honey samples. No statistical relationship was found between colony mortality and pesticide residues. When pesticide residues from all matrices were pooled together, a mixed model analysis did not show a significant relationship between the presence of pesticide residues and the abundance of brood and adults, and no statistical relationship was found between colony mortality and pesticide residues. Thus, although certain pesticide residues were detected in apicultural matrices and occasionally with another pesticide residual, more work is needed to determine the role these residues play in affecting colony health.

An assessment of honeybee colony matrices, Apis mellifera (Hymenoptera: Apidae) to monitor pesticide presence in continental France

The frequency of occurrence and relative concentration of 44 pesticides in apicultural (Apis mellifera) matrices collected from five French locations (24 apiaries) were assessed from 2002 to 2005. The number and nature of the pesticides investigated varied with the matrices examined-living honeybees, pollen loads, honey, and beeswax. Pollen loads and beeswax had the highest frequency of pesticide occurrence among the apiary matrices examined in the present study, whereas honey samples had the lowest. The imidacloprid group and the fipronil group were detected in sufficient amounts in all matrices to allow statistical comparisons. Some seasonal variation was shown when residues were identified in pollen loads. Given the results (highest frequency of presence) and practical aspects (easy to collect; matrix with no turnover, unlike with bees that are naturally renewed), pollen loads were the best matrix for assessing the presence of pesticide residues in the environment in our given conditions.

Colony Collapse Disorder in context

Although most of humanity relies upon foods that do not require animal pollination 1, production of 39 of the worlds 57 most important monoculture crops still benefits from this ecosystem service 2. Western honey bees (Apis mellifera) are undoubtedly the single-most valuable animal pollinators to agriculture because they can be easily maintained and transported to pollinator-dependent crops. Yet, despite an almost 50% increase in world honey bee stocks over the last century, beekeepers have not kept pace with the >300% increase in pollinator-dependent crops 3. This has led to great uncertainty surrounding the recent large-scale die-offs of honey bees around the world, and has sparked enormous interest from both scientists and the general public.

Standard methods for toxicology research in Apis mellifera

Summary Modern agriculture often involves the use of pesticides to protect crops. These substances are harmful to target organisms (pests and pathogens). Nevertheless, they can also damage non-target animals, such as pollinators and entomophagous arthropods. It is obvious that the undesirable side effects of pesticides on the environment should be reduced to a minimum. Western honey bees (Apis mellifera) are very important organisms from an agricultural perspective and are vulnerable to pesticide-induced impacts. They contribute actively to the pollination of cultivated crops and wild vegetation, making food production possible. Of course, since Apis mellifera occupies the same ecological niche as many other species of pollinators, the loss of honey bees caused by environmental pollutants suggests that other insects may experience a similar outcome. Because pesticides can harm honey bees and other pollinators, it is important to register pesticides that are as selective as possible. In this manuscript, we describe a selection of methods used for studying pesticide toxicity/selectiveness towards Apis mellifera. These methods may be used in risk assessment schemes and in scientific research aimed to explain acute and chronic effects of any target compound on Apis mellifera.

Detection of Aethina tumida Murray (Coleoptera: Nitidulidae.) in Italy: outbreaks and early reaction measures

Summary The first detection of Aethina tumida Murray (the small hive beetle) in Italy occurred on 5 September 2014. Three nuclei containing honey bees (Apis mellifera) and located in a clementine (citrus) orchard near an important international harbour in the Calabria region (southern Italy) were heavily infested with adult and larval A. tumida. A. tumida infestation is a notifiable disease of honey bees in the European Union as well as an OIE listed disease. To prevent any A. tumida introduction, the importation of honey bees is regulated strictly in the European Union (Commission Regulation (EU) No. 206/2010). Early reaction measures adopted in Italy require that beekeepers must notify A. tumida discovery to the local veterinary services and cannot move their colonies. Furthermore, a protection area (20 km radius) and surveillance (100 km radius) zone should be established. The surveillance zone now includes the entire territory of Calabria region. Compulsory visits to all apiaries in the protection zone with the collection of the spatial information by means of a georeferentiation process (georeferentiation can be defined as the process to describe a location relative to the earth, in this context the process consists on the collection of the spatial coordinate of a point that represents the spatial location of the apiaries by means of a GPS device) and colony inspection according to 5% expected prevalence (95% CI) are applied. Destruction of infested apiaries is compulsory and the soil under the infested colonies must be ploughed and treated with pyrethroids. If apiaries in the protection zone are found to be negative, traps are placed. In the surveillance zone, veterinarians visit apiaries that are selected according to a risk analysis (migration in infested areas, honey bee or materials exchange) or randomly and colonies are inspected according to 2% expected prevalence (95% CI). Furthermore, in Italy as well in the rest of Europe, investigations are in progress by competent authorities to make an inventory of all bees and colonies moved from Calabria during 2014. Subsequent controls on colonies should be implemented. People from the honey bee network (beekeepers, veterinarians, beekeeping material producers and distributors) should be aware and informed of the hazard posed by A. tumida to honey bees.

A case control study and a survey on mortalities of honey bee colonies (Apis mellifera) in France during the winter of 2005-6.

Summary Several cases of mortality of honey bee colonies (varying from 38 to 100%) were observed in France during the winter of 2005–6. In order to explain the causes of these mortalities, a case control study was conducted on a limited area, together with a larger survey in 18 other apiaries located in 13 sites over the entire country. Both studies included diagnosis of the main honey bee diseases, assessment of the colony management measures taken by beekeepers and the determination of pesticide residues in apicultural matrices. Pollen analysis was carried out on beebread samples to identify which floral species were used for forage before colony death. Poor Varroa destructor treatments together with Nosema disease and brood diseases were frequent in apiaries with high colony mortalities. The absence of any preventive treatment against V. destructorwas the main risk factor.

A horizon scan of future threats and opportunities for pollinators and pollination

Background. Pollinators, which provide the agriculturally and ecologically essential service of pollination, are under threat at a global scale. Habitat loss and homogenisation, pesticides, parasites and pathogens, invasive species, and climate change have been identified as past and current threats to pollinators. Actions to mitigate these threats, e.g., agri-environment schemes and pesticide-use moratoriums, exist, but have largely been applied post-hoc. However, future sustainability of pollinators and the service they provide requires anticipation of potential threats and opportunities before they occur, enabling timely implementation of policy and practice to prevent, rather than mitigate, further pollinator declines. Methods.Using a horizon scanning approach we identified issues that are likely to impact pollinators, either positively or negatively, over the coming three decades. Results.Our analysis highlights six high priority, and nine secondary issues. High priorities are: (1) corporate control of global agriculture, (2) novel systemic pesticides, (3) novel RNA viruses, (4) the development of new managed pollinators, (5) more frequent heatwaves and drought under climate change, and (6) the potential positive impact of reduced chemical use on pollinators in non-agricultural settings. Discussion. While current pollinator management approaches are largely driven by mitigating past impacts, we present opportunities for pre-emptive practice, legislation, and policy to sustainably manage pollinators for future generations.

A case report of a honey bee colony poisoning incident in France

A case report of a honey bee colony poisoning incident in France Marie-Pierre Chauzat, Anne-Claire Martel, Philippe Blanchard, Marie-Claude Clément, Frank Schurr, Cosette Lair, Magali Ribière, Klaus Wallner, Peter Rosenkranz and Jean-Paul Faucon Agence Française de Sécurité Sanitaire des Aliments, Unit of honey bee Pathology, 105 route des Chappes, BP 111, 06 902 Sophia Antipolis cedex, France. Apicultural State Institute, University of Hohenheim, August-von-Hartmannstrasse 13, D-70599 Stuttgart, Germany.